Characterization of Pseudofusicoccum adansoniae, an Endophytic Fungus Residing in Photosynthetic Root of Tinospora cordifolia, a Medicinal Plant

  • Ashish Mishra
  • Surendra K. Gond
  • Vijay K. Sharma
  • Satish K. Verma
  • Jitendra Kumar
  • Dheeraj K. Singh
  • Anuj Kumar
  • Pooja Pandey
  • R. N. KharwarEmail author
Research Article


An endophytic fungus isolated from photosynthetic root of Tinospora cordifolia Miers. was identified as Pseudofusicoccum adansoniae with 100% 18S ITS rRNA partial gene sequence similarity (accession JX951181). The fungus exhibited amylase, lipase and protease activities and was also able to assimilate the galactose, glucose, fructose, lactose and maltose along with l-arginine and alanine. In addition to siderophore production, the fungus was found to be tolerant against various concentrations of NO2, NO3 and NaCl. The maximum biomass and metabolite yield were observed at pH range 5.4–7.0, whereas yield of secondary metabolites was highest in potato dextrose broth (36.66 ± 0.33 mg/100 ml) extracted through ethyl acetate (EtOAc). Interestingly, metabolites extracted through ethyl acetate from PDB and MEB grown cultures were found to be active against all 8 human bacterial pathogens used in this study. The isolation and characterization of P. adansoniae done in this experiment are the initial steps which pave the way for purified isolation of bioactive compounds and enzymes of immense use.


Antimicrobial compounds Enzyme assay Fungal endophytes Photosynthetic root Pseudofusicoccum adansoniae Salt tolerance Tinospora cordifolia 



The authors are thankful to the Head and Coordinator, CAS, Department of Botany, BHU, Varanasi, India, for providing essential research facilities and technical supports. They also thank CSIR and UGC for Financial Support in the form of JRF and SRFs. Financial assistance to one of the authors from DST, New Delhi (File No. SB/EMEQ-121/2014), is gratefully acknowledged.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Trakunyingcharoen T, Cheewangkoon R, To-anun C (2015) Phylogenetic study of the Botryosphaeriaceae species associated with avocado and par rubber in Thailand. Chiang Mai J Sci 42(1):104–116Google Scholar
  2. 2.
    Denman S, Crous PW, Taylo JE, Kang JC, Pascoc I, Wingfield MJ (2000) An overview of the taxonomic history of Botryosphaeria, and a re-evaluation of its anamorphs based on morphology and ITS rDNA phylogeny. Stud Mycol 45:129–140Google Scholar
  3. 3.
    Slippers B, Wingfield MJ (2007) Botryosphaeriaceae as endophytes and latent pathogens of woody plants: diversity, ecology and impact. Fungal Biol Rev 21:90–106CrossRefGoogle Scholar
  4. 4.
    Sharma R, Kulkarni G, Shouche YS (2013) Pseudofusicoccum adansoniae isolated as an endophyte from Jatropha podagrica: new record for India. Mycotaxon 123:39–45CrossRefGoogle Scholar
  5. 5.
    Verma VC, Gond SK, Kumar A, Kharwar RN, Boulanger A, Strobel G (2011) Endophytic fungal flora of roots and fruits of an Indian neem plant Azadirachta indica A. Juss., and impact of culture media on their isolation. Indian J Microbiol 51:469–476CrossRefGoogle Scholar
  6. 6.
    Li HM, Sullivan R, Moy M, Kobayashi DY, Belanger FC (2004) Expression of a novel endophytic fungal chitinase in the infected host grass. Mycologia 96:526–536CrossRefGoogle Scholar
  7. 7.
    Wang JW, Zheng LP, Tan RX (2006) The Preparation of an elicitor from a fungal endophyte to enhance artemisinin production in hairy root cultures of Artemisia annua L. Chin J Biotechnol 22:829–834Google Scholar
  8. 8.
    Gond SK, Kharwar RN, White JF (2014) Will fungi be the new source of the blockbuster drug taxol? Fungal Biol Rev 28:77–84CrossRefGoogle Scholar
  9. 9.
    Singh SS, Pandey SC, Srivastava S, Gupta VS, Patro B, Ghosh AC (2003) Chemistry and medicinal properties of Tinospora cordifolia (Guduchi). Indian J Pharmacol 35:83–91Google Scholar
  10. 10.
    Petrini O, Sieber TN, Toti L, Viret O (1992) Ecology, metabolite production and substrate utilization in endophytic fungi. Nat Toxins 1:185–196CrossRefGoogle Scholar
  11. 11.
    Schulz B, Guske S, Dammann U, Boyle C (1998) Endophyte–host interaction II. Defining symbiosis of the endophyte–host interaction. Symbiosis 25:213–227Google Scholar
  12. 12.
    Sim HJ, Khoo CH, Lee LH, Cheah YK (2010) Molecular diversity of fungal endophytes isolated from Garcinia mangostana and Garcinia parvifolia. J Microbiol Biotechnol 20:651–658CrossRefGoogle Scholar
  13. 13.
    Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA 5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739CrossRefGoogle Scholar
  14. 14.
    Hankin L, Anagnostakis SL (1975) The use of solid media for detection of enzyme production by fungi. Mycologia 67:597–607CrossRefGoogle Scholar
  15. 15.
    Lingappa Y, Lockwood JL (1962) Chitin media for selective isolation and culture of actinomycetes. Phytopathology 52:317–323Google Scholar
  16. 16.
    Kitancharoen N, Hatai K (1998) Some biological characteristics of fungi isolated from salmonid eggs. Mycocience 39:249–255CrossRefGoogle Scholar
  17. 17.
    Pikovskaya RI (1948) Mobilization of phosphorus in soil connection with the vital activity of some microbial species. Microbiologiya 17:362–370Google Scholar
  18. 18.
    Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:46–56CrossRefGoogle Scholar
  19. 19.
    Gupta A, Gupta VK, Modi DR, Yadava LP (2008) Production and characterization of α-amylase from Aspergillus niger. Biotechnology 7(3):551–556CrossRefGoogle Scholar
  20. 20.
    Cupp-Enyard C (2008) Sigma’s non-specific protease activity assay-casein as a substrate. J Vis Exp 19:899Google Scholar
  21. 21.
    Kharwar RN, Maurya AL, Verma VC, Kumar A, Gond SK, Mishra A (2012) Diversity and antimicrobial activity of endophytic fungal community isolated from medicinal plant Cinnamomum camphora. Proc Natl Acad Sci India (Sect B) 82:557–565CrossRefGoogle Scholar
  22. 22.
    Maria GL, Sridhar KR, Raviraja NS (2005) Antimicrobial and enzyme activity of mangrove endophytic fungi of southwest coast of India. J Agric Technol 1:67–80Google Scholar
  23. 23.
    Suryanarayanan TS, Thirunavukkarasu N, Govindarajulu MB, Gopalan V (2012) Fungal endophytes: an untapped source of biocatalysts. Fungal Divers 54:19–30CrossRefGoogle Scholar
  24. 24.
    Rohrmann S, Molitoris P (1992) Screening of wood-degrading enzymes in marine fungi. Can J Bot 70:2116–2123CrossRefGoogle Scholar
  25. 25.
    Selim KA, El-Beih AA, AbdEl-Rahman TM, El-Diwany AI (2012) Biology of endophytic fungi. Curr Res Environ Appl Mycol 2:31–82CrossRefGoogle Scholar
  26. 26.
    Nayak SS, Gonsalves V, Nazareth SW (2012) Isolation and salt tolerance of halophilic fungi from mangroves and solar salterns in Goa–India. Indian J Geo-Mar Sci 41(2):164–172Google Scholar
  27. 27.
    Baltruschat H, Fodor J, Harrach BD, Niemczyk E, Barna B, Gullner G, Janeczko A, Kogel K, Schäfer P, Schwarczinger I, Zuccaro A, Skoczowski A (2008) Salt tolerance of barley induced by the root endophyte Piriformospora indica is associated with a strong increase in antioxidants. New Phytol 180:501–510CrossRefGoogle Scholar
  28. 28.
    Waller F, Achatz B, Baltruschat H, Fodor J, Becker K, Fischer M, Heier T, Huckelhoven R, Neumann C, von Wettstein D, Franken P, Kogel KH (2005) The endophytic fungus Piriformospora indica reprograms barley to salt-stress tolerance, disease resistance, and higher yield. PNAS J 102:13386–13391CrossRefGoogle Scholar
  29. 29.
    Jackson AM, Whipps JM, Lynch JM (1991) Effect of temperature, pH and water potential on growth of four fungi with disease biocontrol potential. World J Microbiol Biotechnol 7:494–501CrossRefGoogle Scholar
  30. 30.
    Kharwar RN, Verma SK, Mishra A, Gond SK, Sharma VK, Kumar J, Singh DK, Afreen T (2011) Assessment of diversity, distribution and antibacterial activity of endophytic fungi isolated from a medicinal plant Adenocalymma alliaceum Miers. Symbiosis 55:39–46CrossRefGoogle Scholar
  31. 31.
    Verma SK, Gond SK, Mishra A, Sharma VK, Kumar J, Singh DK, Kumar A, Goutam J, Kharwar RN (2014) Impact of environmental variables on the isolation, diversity and antibacterial activity of endophytic fungal communities from Madhuca indica Gmel. at different locations in India. Ann Microbiol 64:721–734CrossRefGoogle Scholar
  32. 32.
    Kharwar RN, Mishra A, Gond SK, Stierle A, Stierle D (2011) Anticancer compounds derived from fungal endophytes: their importance and future challenges. Nat Prod Rep 28:1208–1228CrossRefGoogle Scholar
  33. 33.
    Mishra A, Gond SK, Kumar A, Sharma VK, Verma SK, Kharwar RN, Seiber TN (2012) Season and tissue type affect fungal endophyte communities of the Indian medicinal plant Tinospora cordifolia more strongly than geographic location. Microb Ecol 64:388–398CrossRefGoogle Scholar
  34. 34.
    Hassan SRU, Strobel G, Booth E, Kingston B, Floerchinger C, Sears J (2012) Epigenetic modulation of volatile organic compound formation in the mycodiesel producing endophyte-Hypoxylon sp. CI-4. Microbiology 158:465–473CrossRefGoogle Scholar
  35. 35.
    Sun J, Awakawa T, Noguchi H, Abe I (2012) Induced production of mycotoxins in an endophytic fungus from the medicinal plant Datura stramonium L. Bioorg Med Chem Lett 22:6397–6400CrossRefGoogle Scholar
  36. 36.
    Sharma VK, Kumar J, Singh DK, Mishra A, Verma SK, Gond SK, Kumar A, Singh N, Kharwar RN (2017) Induction of cryptic and bioactive metabolites through natural dietary components in an endophytic fungus Colletotrichum gloeosporioides. Front Microbiol 8:1126CrossRefGoogle Scholar

Copyright information

© The National Academy of Sciences, India 2018

Authors and Affiliations

  • Ashish Mishra
    • 1
  • Surendra K. Gond
    • 2
  • Vijay K. Sharma
    • 1
  • Satish K. Verma
    • 1
  • Jitendra Kumar
    • 1
  • Dheeraj K. Singh
    • 1
  • Anuj Kumar
    • 3
  • Pooja Pandey
    • 1
  • R. N. Kharwar
    • 1
    Email author
  1. 1.Mycopathology and Microbial Technology Laboratory, Department of BotanyBanaras Hindu UniversityVaranasiIndia
  2. 2.Department of Botany, Mahila Maha VidyalayaBanaras Hindu UniversityVaranasiIndia
  3. 3.Department of BotanyBuddha Post Graduate CollegeKushinagarIndia

Personalised recommendations